Detection of cell-surface phosphatidylserine using the fluorogenic probe P-IID.
Methods Enzymol
; 640: 291-307, 2020.
Article
en En
| MEDLINE
| ID: mdl-32560803
ABSTRACT
The fluorogenic probe P-IID enables the detection of cell-surface phosphatidylserine (PS) using both fluorescence imaging and flow cytometry. Here we provide a detailed protocol for the use of P-IID for the qualitative detection of externalized PS in apoptotic cells using confocal microscopy, including the real-time imaging of apoptosis upon drug treatment. We also provide a detailed method for the quantitative analysis of cell death by flow cytometry, using P-IID in conjunction with the nuclear stain propidium iodide. P-IID is superior to commonly used Annexin-V fluorophore conjugates for PS detection as it provides a "turn-on" fluorescence response, displays rapid binding kinetics and can be used at low temperature (4°C), without washing and in the absence of Ca2+ ions.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Fosfatidilserinas
/
Apoptosis
Tipo de estudio:
Diagnostic_studies
/
Guideline
/
Qualitative_research
Idioma:
En
Revista:
Methods Enzymol
Año:
2020
Tipo del documento:
Article
País de afiliación:
Australia